机构：[1]Liver Cancer Institute, Zhongshan Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China.[2]Department of Chemistry and NHC Key Laboratory of Glycoconjugates Research, Fudan University, Shanghai 200032, China.[3]Department of Medical Genetics/Prenatal Diagnostic Center, West China Second University Hospital, Sichuan University, Chengdu 610041, China.[4]Key Laboratory of Birth Defects and Related Diseases of Women and Children, Ministry of Education, Sichuan University, Chengdu 610041, China.[5]Liver Cancer Institute, Zhongshan Hospital and Institutes of Biomedical Sciences and Department of Chemistry and NHC Key Laboratory of Glycoconjugates Research, Fudan University, Shanghai 200032, China[6]Department of Medical Genetics/Prenatal Diagnostic Center, West China Second University Hospital and Key Laboratory of Birth Defects and Related Diseases of Women and Children, Ministry of Education, Sichuan University, Chengdu 610041, China
Quantitative glycosylation analysis serves as an effective tool for detecting changes in glycosylation patterns in cancer and various diseases. However, compared with N-glycans, O-glycans present challenges in both qualitative and quantitative mass spectrometry analysis due to their low abundance, ease of peeling, lack of a universal enzyme, and difficult accessibility. To address this challenge, we developed O-GlycoIsoQuant, a novel O-glycome quantitative approach utilizing superbase release and isotopic Girard's P labeling. This method facilitates rapid and efficient nonreducing β-elimination to dissociate O-glycans from proteins using the organic superbase, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), combined with light and heavy isotopic Girard's reagent P (GP) labeling for relative quantification of O-glycans by mass spectrometry. Employing this method, labeled O-glycans exhibit a double peak with a mass difference of 5 Da, suitable for stable relative quantification. The O-GlycoIsoQuant method is characterized by its high labeling efficiency, excellent reproducibility (CV < 20%), and good linearity (R2 > 0.99), across a dynamic range spanning a 100-fold range. This method was applied to various complex sample types, including human serum, porcine spermatozoa, human saliva, and urinary extracellular vesicles, detecting 33, 39, 49, and 37 O-glycans, respectively, thereby demonstrating its broad applicability.
基金：
National Natural Science
Foundation of China (Grants 22174021, and 82121004),
Shanghai Project (22DZ2291700 and 22142202400).
语种：
外文
PubmedID：
中科院(CAS)分区：
出版当年[2024]版：
无
最新[2023]版：
大类|1 区化学
小类|1 区分析化学
第一作者：
第一作者机构：[1]Liver Cancer Institute, Zhongshan Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China.
共同第一作者：
通讯作者：
通讯机构：[5]Liver Cancer Institute, Zhongshan Hospital and Institutes of Biomedical Sciences and Department of Chemistry and NHC Key Laboratory of Glycoconjugates Research, Fudan University, Shanghai 200032, China[6]Department of Medical Genetics/Prenatal Diagnostic Center, West China Second University Hospital and Key Laboratory of Birth Defects and Related Diseases of Women and Children, Ministry of Education, Sichuan University, Chengdu 610041, China
推荐引用方式(GB/T 7714)：
Li Yueyue,Fu Bin,Li Yang,et al.O-GlycoIsoQuant: A Novel O-Glycome Quantitative Approach through Superbase Release and Isotopic Girard's P Labeling[J].Analytical Chemistry.2024,doi:10.1021/acs.analchem.4c01300.
APA：
Li Yueyue,Fu Bin,Li Yang,Li Chong,Zhai Yujia...&Lu Haojie.(2024).O-GlycoIsoQuant: A Novel O-Glycome Quantitative Approach through Superbase Release and Isotopic Girard's P Labeling.Analytical Chemistry,,
MLA：
Li Yueyue,et al."O-GlycoIsoQuant: A Novel O-Glycome Quantitative Approach through Superbase Release and Isotopic Girard's P Labeling".Analytical Chemistry .(2024)
化学